Field of the Invention
The present invention relates to the use of isolated polypeptides having protease activity in animal feed and detergents. It also relates to the use of isolated nucleic acid sequences encoding the proteases in the recombinant production of isolated polypeptides having protease activity and isolated nucleic acid sequences encoding the proteases. The invention also relates to nucleic acid constructs, vectors, and host cells, including plant and animal cells, comprising the nucleic acid sequences, as well as methods for producing and using the proteases, in particular the use of the proteases in animal feed, and detergents.
Description of the Related Art
Proteases of the S1 group and isolated from Saccharopolyspora are known in the art. A protease from Saccharopolyspora erythrea was disclosed by Oliynyk et al., 2007 in ‘Complete genome sequence of the erythromycin-producing bacterium Saccharopolyspora erythraea NRRL23338’, Nat. Biotechnol. 25:447-453. The complete genone sequence was submitted to EMBL/GenBank under accession number AM420293. The amino acid sequence (uniprot: A4FNQ0) is identical to the sequence of SEQ ID NO: 2 (herein) and the mature amino acid sequence is disclosed in SEQ ID NO: 5.
Other S1 proteases are disclosed in the prior art, such as the protease (Uniprot: C7MV18, SEQ ID NO: 9) described by Pati et al., 2009, ‘Complete genome sequence of Saccharomonospora viridis type strain (P101T)’, Stand. Genomic Sci., 1:141-149 with a sequence identity of 80.7% to SEQ ID NO: 5. Yum et al. have described a protease (Uniprot: Q55353, SEQ ID NO: 10) in ‘Purification and characterization of alkaline serine protease from an alkalophilic Streptomyces sp.’, Biosci. Biotechnol. Biochem., 58:470-474 (1994) with sequence identity of 70.4% to SEQ ID NO: 5.
Lucas et al have submitted a protease from Saccharomonospora xinjiangensis XJ-54 with 79.6% sequence identity to SEQ ID NO: 5 to EMBL/GenBank (Uniprot: 10V8H8, SEQ ID NO: 11) and another protease from Saccharomonospora cyanea NA-134 with 79.0% sequence identity to SEQ ID NO: 5 to EMBL/GenBank (Uniprot: H5XEH4, SEQ ID NO: 12). A further protease from Saccharomonospora paurometabolica YIM 90007 with 76.1% sequence identity to SEQ ID NO: 5 has been submitted to EMBL/GenBank (Uniprot: G4J6Q2, SEQ ID NO: 13).
In the patent literature, one protease from Streptomyces sp., which is identical to Uniprot: Q55353 (SEQ ID NO: 10 herein), was disclosed in WO 2005/052146 (SEQ ID NO: 649, Geneseqp: AEA48820) and another protease was disclosed in WO 2005/052161 (SEQ ID NO: 649, Geneseqp: AEA80317) both with sequence identity of 70.4% to SEQ ID NO: 5 for use in animal feed and detergent compositions. Other known proteases have sequence identities that are about or lower than 70%.
The use of proteases in animal feed to improve digestion of proteins in the feed is known. WO 2009/058679 and US 2009/0111161 relating to a protease from Streptomyces (Streptomyces 1AG3 protease) having an identity to the presently indicated protease of SEQ ID NO: 2 of 66%, mention the use of the protease in animal feed. WO 95/28850 discloses the combination of a phytase and one or more microbial proteolytic enzymes to improve the solubility of vegetable proteins. WO 01/58275 discloses the use of acid stable proteases of the subtilisin family in animal feed. WO 01/58276 discloses the use in animal feed of acid-stable proteases related to the protease derived from Nocardiopsis sp. NRRL 18262 (the 10R protease), as well as a protease derived from Nocardiopsis alba DSM 14010. WO 04/072221, WO 04/111220, WO 04/111223, WO 05/035747, and WO 05/123911 disclose proteases related to the 10R protease and their use in animal feed. Also, WO 04/072279 discloses the use of other proteases in animal feed.
WO 04/034776 discloses the use of a subtilisin/keratinase, PWD-1 from B. licheniformis in the feed of poultry. WO 04/077960 discloses a method of increasing digestibility of forage or grain in ruminants by applying a bacterial or fungal protease.
Commercial products comprising a protease and marketed for use in animal feed include RONOZYME® ProAct (DSM NP/Novozymes), Axtra® (Danisco), Avizyme® (Danisco), Porzyme® (Danisco), Allzyme™ (Alltech), Versazyme® (BioResources, Int.), Poultrygrow™ (Jefo) and Cibenza® DP100 (Novus).